High-dose HOOK effect in urinary DcR2 assay in patients with chronic kidney disease.
High-dose HOOK effect in urinary DcR2 assay in patients with chronic kidney disease.
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DOI：
10.1016/j.clinbiochem.2018.06.001
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发表时间：
2018-08
影响因子：
2.8
通讯作者：
Jia Chen;Kehong Chen;Li-ming Wang;Wei-Wei Zhang-Wei;Lei Feng;Huanzi Dai;Ya-ni He
中科院分区：
文献类型：
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作者：
Jia Chen;Kehong Chen;Li-ming Wang;Wei-Wei Zhang-Wei;Lei Feng;Huanzi Dai;Ya-ni He
尿DcR 2（uDcR 2）是早期检测慢性肾脏病（CKD）患者肾小管间质损伤（TII）的生物标志物，但酶联免疫吸附试验（ELISA）检测时，由于高剂量的钩状效应，可能导致检测结果假性偏低甚至阴性。本研究旨在探讨ELISA检测中是否存在高剂量钩状效应，并探索降低这种效应的可能方法。方法收集72例CKD患者，根据TII评分分为4组。使用ELISA试剂盒在未稀释和连续稀释（2倍、4倍、8倍和16倍稀释）的尿液中测量uDcR 2。将测定结果标准化为尿肌酐。我们评估了不同稀释度的uDcR 2/cre水平与肾脏组织学参数之间的相关性。结果CKD患者尿中uDcR 2/cre水平显著高于对照组，尿中uDcR 2/cre水平与TII的相关性显著高于对照组，差异有统计学意义（P < 0. 05）。然而，较高的TII评分并没有产生较高水平的uDcR 2/cre在未稀释的尿液。连续稀释后，uDcR 2/cre水平在4倍稀释时最高。不同稀释度的uDcR 2/cre水平与TII评分之间呈正相关，相关系数最高，AUC最大，在4倍稀释度时观察到。ConclusionsThe high-dose hook effect was apparent during ELISA testing of uDcR 2 in CKD patients，yet dilution of the urine samples needed this effect.然而，使用尿的四倍稀释用于uDcR 2/cre测试可以消除高剂量钩状效应，并且可以有效地监测CKD患者中TII的严重程度。
BackgroundUrinary DcR2 (uDcR2) is a biomarker for the early detection the tubulointerstitial injury (TII) in patients with chronic kidney disease (CKD), but the high-dose hook effect may lead to falsely low or even negative results when using an enzyme-linked immunosorbent assay (ELISA). This study aimed to investigate if the high-dose hook effect exists with ELISA testing, and to uncover a potential approach for reducing this effect.Methods72 CKD patients were recruited and categorized into four groups based on TII scores. uDcR2 was measured in undiluted and serially diluted (two-, four-, eight- and 16-fold dilutions) urine using an ELISA kit. The results from the assay were normalized to urinary creatinine. We evaluated the correlation between uDcR2/cre levels at different dilutions and renal histological parameters. Receiver operating characteristic (ROC) curves were generated to examine the value of uDcR2/cre for predicting TII.ResultsuDcR2/cre levels in the undiluted urine were significantly higher in patients with CKD than those in the control. However, higher TII scores did not yield higher levels of uDcR2/cre in the undiluted urine. After serial dilution, uDcR2/cre levels were highest with the four-fold dilution. A positive correlation was found between uDcR2/cre levels at different dilutions and TII scores, with the highest correlation coefficient and the largest AUC being observed at the four-fold dilution.ConclusionsThe high-dose hook effect was apparent during ELISA testing of uDcR2 in CKD patients, yet dilution of the urine samples neutralized this effect. However, the use of a four-fold dilution of urine for uDcR2/cre testing may eliminate the high-dose hook effect and make it possible to effectively monitor the severity of TII in CKD patients.